Patent US5728528A successfully refreshed from Google Patents.
US5728528A
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US
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Universal spacer/energy transfer dyes
Assignee: University of California
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Filing: 1995-09-20
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Granted: 1998-03-17
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Expires: 2015-09-20
Abstract
Abstract Fluorescent labels are provided employing energy absorber/donor components and energy acceptor/fluorescer components joined together by a spacer which comprises sugar phosphate monomeric units, particularly ribosyl phosphate monomeric units, where the sugar groups are free of active hydroxyl groups. Particularly, an energy transfer component is substituted at the 5' position of the spacer chain, while the other energy transfer component is substituted at the 1' position of the 3' terminal ribosyl group of the label forming an ET cassette for linking to a nucleic acid sequence with any compositions. By employing combinations of ET components, with a common energy absorber/donor and different fluorescers, one can provide for families of labels which can be tagged to any target molecules and which can be excited at a single wavelength and fluoresce at different wavelengths with large Stokes shifts. The compositions find particular application in sequencing.
Inventors
- 👤 Richard A. Mathies
- 👤 Alexander N. Glazer
- 👤 Jingyue Ju
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-29 13:31
CPC Classifications
C; C07; C07H; C07H21/00; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6813; C12Q1/6816; C12Q1/6818; Y; Y10; Y10S; Y10S435/00; Y10S435/81
Claims Analysis
Total Claims: 26
Claim 1
Independent Claim
104 words
In a method for identifying different nucleic acid sequences using a combination of fluorescent labels in an electrophoresis separation, said fluorescent labels fluorescing at different wavelengths and each different label joined to a different nucleic acid sequence, the improvement which comprises: said labels comprising an energy donor component absorbing at a common wavelength, an acceptor component absorbing energy transferred by said energy donor component, where the acceptor for each label emits at a different wavelength, and said energy donor component and acceptor component linked by a backbone comprising sugar phosphate ester monomer linkages with the 1'-hydroxyl group being replaced with a hydrogen or etherified.
Claim 2
Dependent Claim
104 words
In a method for identifying different nucleic acid sequences using a combination of fluorescent labels in an electrophoresis separation, said fluorescent labels fluorescing at different wavelengths and each different label joined to a different nucleic acid sequence, the improvement which comprises: said labels comprising an energy donor component absorbing at a common wavelength, an acceptor component absorbing energy transferred by said energy donor component, where the acceptor for each label emits at a different wavelength, and said energy donor component and acceptor component linked by a backbone comprising sugar phosphate ester monomer linkages with the 1'-hydroxyl group being replaced with a hydrogen or etherified.
Claim 3
Dependent Claim
16 words
A method according to claim 1, wherein said sugar phosphate ester linkages are 1',2'-dideoxyribosyl-3', 5'-phosphate linkages.
Claim 4
Dependent Claim
16 words
A method according to claim 2, wherein said backbone comprises 3 to 15 phosphate ester monomers.
Claim 5
Dependent Claim
25 words
A method according to claim 3, wherein said backbone has a 5' hydroxyl terminus and said energy donor component is linked at said 5' terminus.
Claim 6
Dependent Claim
32 words
A method according to claim 3, wherein said backbone has a 3' terminus comprising a linking functionality at the 1'-position of said 1', 2'-dideoxyribosyl group to which said acceptor is covalently bonded.
Claim 7
Dependent Claim
131 words
In a method for identifying different nucleic acid sequences using a combination of fluorescent labels in an electrophoresis separation, said fluorescent labels linked to the appropriate primers and fluorescing at different wavelengths and each different label joined to a different nucleic acid sequence, the improvement which comprises: said labels comprising an energy donor component absorbing at a common wavelength, an acceptor component absorbing energy transferred by said donor component, where the acceptor for each label emits at a different wavelength, and said energy donor component and acceptor component are linked by a backbone comprising from 3 to 15 1',2'-dideoxyribosyl 3',5'-phosphate ester monomer linkages, said energy donor component covalently linked at the 5' terminus of said backbone and said acceptor component linked to the 1'-position of the 3'-terminal 1',2'-dideoxyribose of said backbone.
Claim 8
Dependent Claim
131 words
In a method for identifying different nucleic acid sequences using a combination of fluorescent labels in an electrophoresis separation, said fluorescent labels linked to the appropriate primers and fluorescing at different wavelengths and each different label joined to a different nucleic acid sequence, the improvement which comprises: said labels comprising an energy donor component absorbing at a common wavelength, an acceptor component absorbing energy transferred by said donor component, where the acceptor for each label emits at a different wavelength, and said energy donor component and acceptor component are linked by a backbone comprising from 3 to 15 1',2'-dideoxyribosyl 3',5'-phosphate ester monomer linkages, said energy donor component covalently linked at the 5' terminus of said backbone and said acceptor component linked to the 1'-position of the 3'-terminal 1',2'-dideoxyribose of said backbone.
Claim 9
Dependent Claim
21 words
A method according to claim 6, wherein said backbone comprises from 5 to 8 1', 2'-dideoxyribosyl 3', 5'-phosphate ester monomer linkages.
Claim 10
Dependent Claim
13 words
A method according to claim 6, wherein said donor component is a fluorescein.
Claim 11
Dependent Claim
14 words
A method according to claim 6, wherein said acceptor is linked through a uridine.
Claim 12
Dependent Claim
26 words
A method according to claim 6, wherein said acceptor is linked through a carboxy group bonded to the 1'-position at the 3'-terminus of the sugar monomer.
Claim 13
Dependent Claim
14 words
A method according to claim 6, wherein at least one fluorescer is a rhodamine.
Claim 14
Dependent Claim
147 words
In a method for identifying different nucleic acid sequences using a combination of fluorescent labels in an electrophoresis separation for determining a nucleic acid sequence, where each of said labels is associated with the presence of a different nucleotide, said fluorescent labels fluorescing at different wavelengths and each different label joined to a different nucleic acid sequence, the improvement which comprises: said labels comprising an energy donor component absorbing at a common wavelength, an acceptor component absorbing energy from said energy donor component, where the acceptor for each label emits at a different wavelength, and said energy donor component and acceptor component linked by a backbone comprising from 3 to 9 1', 2'-dideoxyribosyl 3', 5'-phosphate ester monomer linkages, said energy donor component covalently linked at the 5' terminus of said backbone and said acceptor component linked to the 1'-position of the 3'-terminal 1', 2'-dideoxyribose of said backbone.
Claim 15
Dependent Claim
147 words
In a method for identifying different nucleic acid sequences using a combination of fluorescent labels in an electrophoresis separation for determining a nucleic acid sequence, where each of said labels is associated with the presence of a different nucleotide, said fluorescent labels fluorescing at different wavelengths and each different label joined to a different nucleic acid sequence, the improvement which comprises: said labels comprising an energy donor component absorbing at a common wavelength, an acceptor component absorbing energy from said energy donor component, where the acceptor for each label emits at a different wavelength, and said energy donor component and acceptor component linked by a backbone comprising from 3 to 9 1', 2'-dideoxyribosyl 3', 5'-phosphate ester monomer linkages, said energy donor component covalently linked at the 5' terminus of said backbone and said acceptor component linked to the 1'-position of the 3'-terminal 1', 2'-dideoxyribose of said backbone.
Claim 16
Dependent Claim
42 words
A compound comprising an energy donor component and an acceptor component absorbing energy transferred by said energy donor component, said components linked by a backbone comprising sugar phosphate ester monomer linkages with the 1'-hydroxyl group being replaced by a hydrogen or etherified.
Claim 17
Dependent Claim
42 words
A compound comprising an energy donor component and an acceptor component absorbing energy transferred by said energy donor component, said components linked by a backbone comprising sugar phosphate ester monomer linkages with the 1'-hydroxyl group being replaced by a hydrogen or etherified.
Claim 18
Dependent Claim
21 words
A compound according to claim 13, wherein said backbone comprises from 3 to 15 1', 2'-dideoxyribosyl 3', 5'-phosphate ester monomer linkages.
Claim 19
Dependent Claim
42 words
A compound according to claim 14, wherein said energy donor component is covalently bonded at the 5' backbone terminus and said acceptor is covalently bonded to the backbone 3'-terminal of 1', 2'-dideoxyribosyl 3', 5'-phosphate ester at the 1'-position through a linking group.
Claim 20
Dependent Claim
22 words
A compound according to claim 15, wherein the 3'-terminus of said backbone is an hydroxyl, protected hydroxyl, phosphoramidite, phosphite, nucleotide or oligonucleotide.
Claim 21
Dependent Claim
44 words
A compound of the formula 5'-fluorescein substituted poly-1', 2'-dideoxyribose-3', 5'-phosphate, having from 3 to 15 1', 2'-dideoxyribose-3', 5'-phosphate monomeric units, and substituted through a linking group at the 1'-position of the 3'-terminal 1', 2'-dideoxyribose-3', 5' -phosphate monomeric unit with a fluorescein or a rhodamine.
Claim 22
Dependent Claim
44 words
A compound of the formula 5'-fluorescein substituted poly-1', 2'-dideoxyribose-3', 5'-phosphate, having from 3 to 15 1', 2'-dideoxyribose-3', 5'-phosphate monomeric units, and substituted through a linking group at the 1'-position of the 3'-terminal 1', 2'-dideoxyribose-3', 5' -phosphate monomeric unit with a fluorescein or a rhodamine.
Claim 23
Dependent Claim
22 words
A compound according to claim 17, wherein the 3'-terminus of said backbone is an hydroxyl, protected hydroxyl, phosphoramidite, phosphite, nucleotide or oligonucleotide.
Claim 24
Dependent Claim
16 words
A compound according to claim 17, wherein said linking group is a carboxyl or uridyl group.
Claim 25
Dependent Claim
21 words
A kit comprising at least two compounds according to claim 13 differing in said acceptor and having the same energy donor.
Claim 26
Dependent Claim
21 words
A kit comprising at least two compounds according to claim 15 differing in said acceptor and having the same energy donor.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 2001-03-16 | 2001-09-15 | 2002-03-17 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2005-03-16 | 2005-09-15 | 2006-03-17 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2009-03-16 | 2009-09-15 | 2010-03-17 | $7700.00 | Window Closed / Lapsed |